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cDNA cloning and 1.75 Å crystal structure determination of PPL2, an endochitinase and N ‐acetylglucosamine‐binding hemagglutinin from Parkia platycephala seeds
Author(s) -
Cavada Benildo S.,
Moreno Frederico B. B.,
da Rocha Bruno A. M.,
de Azevedo Walter F.,
Castellón Rolando E. R.,
Goersch Georg V.,
Nagano Celso S.,
de Souza Emmanuel P.,
Nascimento Kyria S.,
RadisBaptista Gandhi,
Delatorre Plínio,
Leroy Yves,
Toyama Marcos H.,
Pinto Vicente P. T.,
Sampaio Alexandre H.,
Barettino Domingo,
Debray Henri,
Calvete Juan J.,
Sanz Libia
Publication year - 2006
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2006.05400.x
Subject(s) - lectin , biology , biochemistry
Parkia platycephala lectin 2 was purified from Parkia platycephala (Leguminosae, Mimosoideae) seeds by affinity chromatography and RP‐HPLC. Equilibrium sedimentation and MS showed that Parkia platycephala lectin 2 is a nonglycosylated monomeric protein of molecular mass 29 407 ± 15 Da, which contains six cysteine residues engaged in the formation of three intramolecular disulfide bonds. Parkia platycephala lectin 2 agglutinated rabbit erythrocytes, and this activity was specifically inhibited by N ‐acetylglucosamine. In addition, Parkia platycephala lectin 2 hydrolyzed β(1–4) glycosidic bonds linking 2‐acetoamido‐2‐deoxy‐β‐ d ‐glucopyranose units in chitin. The full‐length amino acid sequence of Parkia platycephala lectin 2, determined by N‐terminal sequencing and cDNA cloning, and its three‐dimensional structure, established by X‐ray crystallography at 1.75 Å resolution, showed that Parkia platycephala lectin 2 is homologous to endochitinases of the glycosyl hydrolase family 18, which share the (βα) 8 barrel topology harboring the catalytic residues Asp125, Glu127, and Tyr182.